Detection Of Herpes Simplex Virus DNA Research Articles

A cost-effective label-free biosensor for rapid detection of multiple viral respiratory infections base on liquid crystals: Fabrication and modeling

The high transmission rate and incurability of recurrent Herpes Simplex Virus (HSV) infection require an accurate and fast detection approach for its efficient prevention. In an attempt to find a novel sensitive and simple optical diagnostic technique which enables on-spot testing, here we developed label-free biosensors based on E7 nematic liquid crystals (LCs) for specific detection of HSV DNA. For this purpose, homeotropically oriented LCs as smart materials are spread on the textile grid to make their both upper and lower surfaces be in contact with ambient air. HSV DNA of concentration between 1.4 nM and 5.5 nM is added to the platform, and DNA elution buffer is used as a control. Then, the reorientation of LCs occurs due to the coupling of electrical dipoles between LCs and DNA molecules. Creating a unique pattern that is projected onto CCD by putting the sample in a cross-polarizer and using white light. To model the LC's director distribution, LC's free energy equation is solved using the variational method and finite difference method (FDM). Overall, our LC-based sensor is a user-friendly, cost-effective, highly sensitive, and fast way for specific detection of HSV without the need for experts and expensive equipment.

An Enigmatic Case of a Febrile Infant With Seizures

Herpes simplex virus (HSV) encephalitis is one of the most common viral infections in infants associated with high morbidity and mortality rates despite available antiviral therapy. For symptomatic infants, starting empiric therapy with acyclovir can prevent serious neurological sequelae while awaiting results from diagnostic studies. The gold standard of diagnosis remains to be the detection of HSV DNA via polymerase chain reaction (PCR) from cerebrospinal fluid (CSF). However, due to the low viral load in the initial stages of infection, even the gold standard test may not detect active infection. We present a case of an eight-month-old child who presented with fever and seizures and had negative HSV DNA PCR from initial CSF studies. Ongoing fever and recurrent seizures prompted an MRI which was suggestive of meningoencephalitis, HSV DNA PCR from repeat CSF sample resulted positive. This case emphasizes the importance of keen clinical judgment and the caution required when deciding to stop empiric therapy when the clinical suspicion for HSV encephalitis remains high.

Herpes simplex virus DNA testing by a loop-mediated isothermal amplification method for accurate clinical diagnosis and detection of mucosal viral shedding.

Herpes simplex virus (HSV) periodically forms characteristic blisters in the perioral and genital areas in a subset of people. Because of the lack of accurate tests for this common virus, various types of perioral/anogenital lesions are often misdiagnosed as herpes. Also, though asymptomatic HSV-positive people shed virus, the precise time course of symptoms and viral shedding is unclear. The loop-mediated isothermal amplification (LAMP) method amplifies target DNA sequences without thermal cycles, simpler and faster than polymerase chain reaction (PCR). To investigate clinico-laboratorial correlation and whether HSV can be detected in the oral cavity during symptom occurrence, we collected 445 specimens from 211 patients who visited our clinic with suspected herpetic lesions or non-symptomatic volunteers. DNA was extracted from swabs simultaneously taken from lesions (n=219) and seemingly asymptomatic oral mucosa (n=226). HSV-1 and HSV-2 DNA sequences were amplified by LAMP and validated by quantitative real-time PCR. The LAMP method detected HSV DNA almost as sensitively (97%) as PCR. Positivity for HSV DNA was found in 54% (40/74) of specimens from the perioral/oral area. Review of clinical images of recurrent herpes labialis revealed that HSV DNA was detected only from lesions located on the perioral skin and/or the dry, vermillion part of the lip; no HSV DNA was found in immunocompetent patients with lesions confined to the oral mucosa except primary infection. This observation may be an important principle for clinical diagnosis of recurrent herpes. HSV was detected in the oral mucosa in 2.7% (6/226) of samples; all of these patients had either primary infection or were immunosuppressed. Virus shedding in the mucosa was apparently tightly regulated by the immune system. Patients with suppressed or no immunity (naïve cases) did shed virus in the mucosa. LAMP is a simple method to reliably distinguish recurrent/primary herpes from other conditions.

Rapid diagnostics of genital herpes by loop-mediated isothermal amplification method with fluorescent detection

Introduction. Due to the high clinical significance of herpesvirus diseases, the searching of fast and effective methods for their diagnosis remains relevant.The aim of the study was to evaluate the diagnostic efficiency of the loop-mediated isothermal amplification of DNA with real-time fluorescent detection (RT-LAMP) with SYTO-82 dye on a model of herpes simplex virus (HSV) infection.Materials and methods. A total of 44 urogenital swabs containing type 1 and type 2 HSV DNA and 43 swabs without HSV DNA, including 33 samples containing the DNA of cytomegalovirus, Epstein-Barr virus and herpesvirus type 6, were studied. For RT-LAMP, Bst 2.0 WarmStart DNA polymerase, SYTO-82 dye, LAMP primers were used.Results. The high efficiency of HSV DNA detection in the RT-LAMP reaction with SYTO-82 dye was shown. RT-LAMP in optimal conditions allowed to reduce reaction time for 2-3 times compared to real-time PCR (to 35 minutes). Analytical sensitivity of HSV type 1 and 2 detection in RT-LAMP was 103 copies of DNA/ml. The diagnostic sensitivity and specificity of the RT-LAMP diagnosis of HSV infection were 96% and 100%, respectively.Discussion. RT-LAMP method has a high sensitivity and specificity comparable to RTPCR, while the risk of false positive results obtaining is minimal.Conclusion. Thus, the reaction of RT-LAMP with SYTO-82 dye allows quickly, with high sensitivity and specificity to detect HSV DNA in clinical material and can be considered as a promising point-of-care testing method.

64-Year-Old Man With Subacute Altered Mental Status and Headache

A 64-year-old man from Iowa with a medical history of sarcoidosis, managed with prednisone, 20 mg/d, for the past 5 months, presented to the emergency department with gradual-onset, progressive altered mental status and headaches of 1 month’s duration. Previously, he was cognitively intact according to his daughter. He had become unable to perform normal daily tasks such as unlocking doors and holding normal conversations. Additionally, he reported a generalized, throbbing, constant headache associated with intermittent blurry vision. About 1 week before presentation, he also experienced severe fatigue and lethargy in addition to a few falls. He had no fever, weight loss, syncope, seizures, or neck stiffness, no exposure to birds or their droppings, no history of travel to the American Southwest or elsewhere, and no sick contacts. On presentation, his vital signs revealed a temperature of 36.6°C; blood pressure, 148/103 mm Hg; heart rate, 104 beats/min; and oxygen saturation, 98% while breathing room air. He appeared intermittently confused but had no meningismus, papilledema, or focal neurologic deficits on physical examination. Initial laboratory evaluation revealed the following (reference ranges provided parenthetically): hemoglobin, 16.8 g/dL (13.2-16.6 g/dL); leukocytes, 10.2 × 109/L (3.4-9.6 × 109/L); platelets, 261 × 109/L (135-317 × 109/L); electrolytes within normal limits; thyrotropin, 1.8 mIU/L (0.3-4.2 mIU/L); and prothrombin time, 13.3 seconds (9.4-12.5 seconds). Urinalysis results were normal. Acetaminophen and salicylate levels were undetectable. Computed tomography of the head revealed no bleeding, masses, or other changes that would explain the clinical picture.1. Based on this patient’s presentation, which one of the following diagnostic tests should be performed next?a.Magnetic resonance imaging (MRI) of the brainb.Electroencephalographyc.Lumbar puncture (LP)d.Angiotensin-converting enzyme measuremente.Electromyography Magnetic resonance imaging of the brain is a good test to evaluate changes within the brain parenchyma and can also help in the diagnosis of herpes simplex virus (HSV) encephalitis. However, it is expensive, time consuming, and not readily available in all practice settings. The criterion standard for diagnosis of HSV encephalitis is the detection of HSV DNA in the cerebrospinal fluid (CSF) by polymerase chain reaction (PCR). Although brain MRI might be needed later on, it is not the best next step in this patient’s situation. Electroencephalography could help evaluate for seizure activity or epilepsy, but the patient has no history suggestive of such disorders at this presentation. The best diagnostic step in evaluating a patient, especially if immunocompromised, with subacute altered mental status, headaches, and no focal neurologic deficit is LP. Meningitis and encephalitis must be high on the differential diagnosis and would require CSF analysis for diagnosis. Lumbar puncture can be performed easily by many physicians in different practice settings, has a low complication risk, and is less expensive than MRI. The differential diagnosis may include neurosarcoidosis as well, but it usually presents with cranial nerve palsies and less commonly with headaches.1Zajicek J.P. Scolding N.J. Foster O. et al.Central nervous system sarcoidosis—diagnosis and management.QJM. 1999; 92: 103-117Crossref PubMed Scopus (600) Google Scholar Furthermore, angiotensin-converting enzyme levels have low sensitivity and specificity in diagnosing sarcoidosis.2Ungprasert P. Carmona E.M. Crowson C.S. Matteson E.L. Diagnostic utility of angiotensin-converting enzyme in sarcoidosis: a population-based study.Lung. 2016; 194: 91-95Crossref PubMed Scopus (100) Google Scholar Electromyography may be useful for evaluating proximal muscle weakness or the presence of peripheral neuropathy, which are absent in this patient. Lumbar puncture yielded clear CSF. Initial CSF analysis revealed the following (reference ranges provided parenthetically): opening pressure, 25 cm H2O (7-18 cm H2O); total protein, 107 mg/dL (0-35 mg/dL); glucose, 35 mg/dL, with a serum glucose level of 102 mg/dL (CSF glucose should be 60% of serum glucose when drawn concurrently); total nucleated cells, 34/μL (0-5/μL); erythrocytes, 0% (0%); neutrophils, 3% (0%-6%); lymphocytes, 69% (40%-80%); and monocytes, 28% (15%-45%).2.Given the overall presentation so far, which one of the following additional CSF tests would provide the highest diagnostic yield?a.Streptococcus pneumoniae PCRb.Neisseria meningitidis PCRc.Enterovirus PCRd.Coccidioidomycosis antigen teste.Cryptococcal antigen test Streptococcus pneumoniae and N meningitides are common pathogens in acute bacterial meningitis, which presents with fever, nuchal rigidity, and characteristic CSF findings of pleocytosis (cell count over 100), low glucose level, and high protein value.3Durand M.L. Calderwood S.B. Weber D.J. et al.Acute bacterial meningitis in adults: a review of 493 episodes.N Engl J Med. 1993; 328: 21-28Crossref PubMed Scopus (1160) Google Scholar This scenario does not fit with our patient’s subacute/chronic disease course nor with results of his CSF analysis. Enterovirus causes aseptic meningitis, characterized by a rapid onset (within hours) as opposed to the month-long course our patient experienced. Additionally, the cost and lack of actionable treatment, combined with generally good outcomes, makes the clinical utility of enterovirus PCR low in many cases.4Bahr N.C. Boulware D.R. Methods of rapid diagnosis for the etiology of meningitis in adults.Biomark Med. 2014; 8: 1085-1103Crossref PubMed Google Scholar Coccidioidal meningitis can present with subacute headache and mental status changes, but it is primarily found in the southwestern United States. Our patient is from Iowa and had no recent travel. Cryptococcal antigen testing would provide the highest diagnostic yield in this patient. First, his clinical course is consistent with subacute meningoencephalitis, characterized by a subacute headache and gradual mental status changes over 2 to 4 weeks, which is seen with cryptococcal meningitis (CM).5Pappas P.G. Perfect J.R. Cloud G.A. et al.Cryptococcosis in human immunodeficiency virus-negative patients in the era of effective azole therapy.Clin Infect Dis. 2001; 33: 690-699Crossref PubMed Scopus (490) Google Scholar Second, his initial CSF findings of mildly elevated nucleated cell count with a lymphocyte predominance, low glucose concentration, and high protein level are suggestive of a fungal or mycobacterial infection, the latter being less common. When CM is suspected, sending the CSF for cryptococcal antigen testing is the appropriate next step. It can be analyzed quickly and has a sensitivity of 93% to 100% and a specificity of 93% to 98%.6Tanner D.C. Weinstein M.P. Fedorciw B. Joho K.L. Thorpe J.J. Reller L. Comparison of commercial kits for detection of cryptococcal antigen.J Clin Microbiol. 1994; 32: 1680-1684PubMed Google Scholar, 7Abassi M. Boulware D.R. Rhein J. Cryptococcal meningitis: diagnosis and management update.Curr Trop Med Rep. 2015; 2: 90-99Crossref PubMed Scopus (80) Google Scholar Our patient was admitted to the hospital. His CSF studies were positive for cryptococcal antigen, with a high titer of greater than 1:2560. Cerebrospinal fluid cultures grew Cryptococcus neoformans after 25 days of inoculation. Results of human immunodeficiency virus (HIV) serology and CSF mycobacterial cultures were negative. With a confirmed diagnosis of CM, input from the infectious disease department was sought to provide recommendations for treatment.3.Which one of the following is the preferred induction therapy for CM in this patient?a.Flucytosine for at least 2 weeksb.Liposomal amphotericin B for at least 2 weeksc.Fluconazole for at least 2 weeksd.Liposomal amphotericin B and flucytosine for at least 2 weekse.Liposomal amphotericin B and fluconazole for at least 2 weeks A quick and effective reduction of the yeast burden within the central nervous system (CNS) is the goal of induction therapy. Each one of the first 3 medications—flucytosine, liposomal amphotericin B, and fluconazole—may be active in vitro against cryptococcosis, but monotherapy with any one of them is not recommended given the better and faster fungicidal activity of combination therapy. For patients with CM who are HIV negative and have not had a transplant, the current clinical practice guidelines recommend induction therapy with amphotericin B deoxycholate (0.7-1.0 mg/kg per day intravenously [IV]) and flucytosine (100 mg/kg per day orally in 4 divided doses).8Perfect J.R. Dismukes W.E. Dromer F. et al.Clinical practice guidelines for the management of cryptococcal disease: 2010 update by the Infectious Diseases Society of America.Clin Infect Dis. 2010; 50: 291-322Crossref PubMed Scopus (1768) Google Scholar This combination is associated with improved survival compared with amphotericin B monotherapy. No survival benefit of amphotericin B plus fluconazole was found in a recent randomized trial.9Day J.N. Chau T.T.H. Wolbers M. et al.Combination antifungal therapy for cryptococcal meningitis.N Engl J Med. 2013; 368: 1291-1302Crossref PubMed Scopus (314) Google Scholar Given the higher toxicity with amphotericin B deoxycholate, lipid formulations including liposomal amphotericin B (3-4 mg/kg per day IV) and amphotericin B lipid complex (5 mg/kg per day IV) are often substituted for amphotericin B deoxycholate in the United States. Induction therapy is recommended for at least 4 weeks if there are no neurologic complications. In patients who are at low risk for therapeutic failure, induction therapy can be shortened to only 2 weeks. Such patients include those who have an early diagnosis by history, no uncontrolled underlying disease or immunocompromised state, and excellent clinical response to initial 2-week induction therapy. Conversely, in patients who do exhibit focal neurologic complications, induction therapy can be extended to 6 weeks. Following induction therapy, consolidation therapy with fluconazole (800 mg/d orally) should be given for 8 weeks. After induction and consolidation therapy, patients should receive maintenance therapy with fluconazole (200 mg/d orally) for 6 to 12 months.8Perfect J.R. Dismukes W.E. Dromer F. et al.Clinical practice guidelines for the management of cryptococcal disease: 2010 update by the Infectious Diseases Society of America.Clin Infect Dis. 2010; 50: 291-322Crossref PubMed Scopus (1768) Google Scholar Liposomal amphotericin B and flucytosine were administered, but the patient continued to have morning headaches a few days later.4.Which one of the following is the preferred first step in the management of this patient’s morning headaches?a.Acetazolamideb.Mannitolc.Corticosteroidsd.Serial LPse.Ventriculoperitoneal shunt Our patient has an elevated intracranial pressure (ICP), supported by an opening pressure of 25 cm H2O during his LP. This common complication of CM is responsible for his morning headaches. Acetazolamide, mannitol, and corticosteroids have no proven benefit in this setting, so they are not recommended.8Perfect J.R. Dismukes W.E. Dromer F. et al.Clinical practice guidelines for the management of cryptococcal disease: 2010 update by the Infectious Diseases Society of America.Clin Infect Dis. 2010; 50: 291-322Crossref PubMed Scopus (1768) Google Scholar The preferred first step in management is serial LPs to reduce his opening pressure to a normal of less than 20 cm H2O. Ventriculoperitoneal shunts can be placed only if more conservative measures to control increased ICP have failed.7Abassi M. Boulware D.R. Rhein J. Cryptococcal meningitis: diagnosis and management update.Curr Trop Med Rep. 2015; 2: 90-99Crossref PubMed Scopus (80) Google Scholar, 8Perfect J.R. Dismukes W.E. Dromer F. et al.Clinical practice guidelines for the management of cryptococcal disease: 2010 update by the Infectious Diseases Society of America.Clin Infect Dis. 2010; 50: 291-322Crossref PubMed Scopus (1768) Google Scholar After induction therapy with liposomal amphotericin B and flucytosine and repeated LP, the patient’s symptoms improved dramatically. Morning headaches resolved, and mental status returned to his baseline. He was being prepared for hospital dismissal with appropriate follow-up.5.Which one of the following would be the preferred follow-up after starting induction therapy for CM in this patient?a.Daily measurement of electrolytes and magnesiumb.Weekly measurement of electrolytes and magnesiumc.Daily measurement of serum cryptococcal antigend.Weekly measurement of CSF cryptococcal antigene.Weekly measurement of CD4 and CD3 counts Although induction therapy with liposomal amphotericin B and flucytosine has been found to improve mortality in CM, it is important to recognize the severe adverse effects these medications can have. Amphotericin B deoxycholate in particular is known to be nephrotoxic and causes major electrolyte abnormalities that can be life-threatening. Although lipid formulations of amphotericin B are less nephrotoxic, they still require monitoring of creatinine and electrolyte levels. Potassium and magnesium wasting typically begins after 5 days of amphotericin therapy.7Abassi M. Boulware D.R. Rhein J. Cryptococcal meningitis: diagnosis and management update.Curr Trop Med Rep. 2015; 2: 90-99Crossref PubMed Scopus (80) Google Scholar Flucytosine also has hematologic adverse effects, including leukopenia and thrombocytopenia, that can worsen in acute-on-chronic kidney disease. As such, it is important to obtain daily electrolyte measurements to monitor for and treat these abnormalities. Daily measurement of serum cryptococcal antigen and weekly measurement of CSF cryptococcal antigen are not reliable measures to provide therapeutic monitoring and thus are not routinely recommended for that purpose.10Antinori S. Radice A. Galimberti L. Magni C. Fasan M. Parravicini C. The role of cryptococcal antigen assay in diagnosis and monitoring of cryptococcal meningitis [letter].J Clin Microbiol. 2005; 43: 5828-5829Crossref PubMed Scopus (62) Google Scholar Because CM can be seen in apparently immunocompetent individuals, measurement of CD4 and CD3 counts would not help predict response to therapy. Cryptococcal meningitis is a devastating opportunistic infection caused by the encapsulated yeast C neoformans. It is often considered in HIV-infected patients, but it can occur in patients with impaired cell-mediated immunity from other etiologies.11Lin Y.Y. Shiau S. Fang C.T. Risk factors for invasive Cryptococcus neoformans diseases: a case-control study.PloS One. 2015; 10: e0119090Crossref PubMed Scopus (70) Google Scholar In a US study of more than 300 HIV-negative patients with cryptococcal infection, half had CNS involvement, and of these, 25% had received corticosteroid therapy, 24% had chronic liver, kidney, or lung disease, 16% had a malignancy, 15% had received solid organ transplants, and 30% had no apparent underlying condition.5Pappas P.G. Perfect J.R. Cloud G.A. et al.Cryptococcosis in human immunodeficiency virus-negative patients in the era of effective azole therapy.Clin Infect Dis. 2001; 33: 690-699Crossref PubMed Scopus (490) Google Scholar These findings are important given the considerable morbidity and mortality regardless of HIV status, making early recognition key to expedite management.12Williamson P.R. Jarvis J.N. Panackal A.A. et al.Cryptococcal meningitis: epidemiology, immunology, diagnosis and therapy.Nat Rev Neurol. 2017; 13: 13-24Crossref PubMed Scopus (246) Google Scholar Our patient has 2 risk factors that impair cell-mediated immunity, including sarcoidosis and corticosteroid therapy, which predisposes him to CM. Patients with CM usually present with subacute or chronic meningitis symptoms of a few weeks’ duration. However, these symptoms, such as headache and altered mental status, are not specific. When suspected, LP is the best first step in diagnosing CM. Typically, CSF studies will reveal a high opening pressure, lymphocytic pleocytosis, low glucose concentration, and high protein level. In general, CM should be considered in all cases of lymphocytic meningitis. Cerebrospinal fluid cryptococcal antigen testing has a rapid turnaround time and can establish the diagnosis with high specificity and sensitivity.6Tanner D.C. Weinstein M.P. Fedorciw B. Joho K.L. Thorpe J.J. Reller L. Comparison of commercial kits for detection of cryptococcal antigen.J Clin Microbiol. 1994; 32: 1680-1684PubMed Google Scholar, 7Abassi M. Boulware D.R. Rhein J. Cryptococcal meningitis: diagnosis and management update.Curr Trop Med Rep. 2015; 2: 90-99Crossref PubMed Scopus (80) Google Scholar Serum cryptococcal antigen testing is helpful especially in situations in which CSF assessment is delayed for any reason. Additionally, fungal cultures of blood and CSF specimens can be helpful in the diagnostic work-up. The cryptococcal polysaccharide capsule causes a physical obstruction of the arachnoid villi, which leads to a failure to resorb CSF and in turn elevated ICP.12Williamson P.R. Jarvis J.N. Panackal A.A. et al.Cryptococcal meningitis: epidemiology, immunology, diagnosis and therapy.Nat Rev Neurol. 2017; 13: 13-24Crossref PubMed Scopus (246) Google Scholar Interestingly, the elevation in ICP is positively correlated with an increased number of organisms in the CSF. One feared complication of intracranial hypertension is cerebral edema. Common symptoms and signs include headaches, vomiting, visual changes, papilledema, abducens nerve palsy, altered mental status, and ultimately coma. Increased ICP in patients with CM should be managed aggressively to decrease morbidity and mortality and is usually achieved using serial LPs.8Perfect J.R. Dismukes W.E. Dromer F. et al.Clinical practice guidelines for the management of cryptococcal disease: 2010 update by the Infectious Diseases Society of America.Clin Infect Dis. 2010; 50: 291-322Crossref PubMed Scopus (1768) Google Scholar Once the diagnosis of CM is confirmed, it is important to promptly initiate appropriate antifungal therapy. Although fluconazole monotherapy can be used for mild cryptococcal disease not involving the CNS, treatment of CM is lengthy and arduous and includes 3 stages: induction, consolidation, and maintenance. Induction therapy consists of deoxycholate or lipid formulations of amphotericin B plus flucytosine for 4 to 6 weeks. Subsequently, consolidation therapy should be initiated with fluconazole (800 mg/d orally) for 8 weeks. Following that, maintenance therapy with fluconazole (200 mg/d orally) should be given for 6 to 12 months.8Perfect J.R. Dismukes W.E. Dromer F. et al.Clinical practice guidelines for the management of cryptococcal disease: 2010 update by the Infectious Diseases Society of America.Clin Infect Dis. 2010; 50: 291-322Crossref PubMed Scopus (1768) Google Scholar Of note, all these medications can have serious adverse effects. Amphotericin B products in particular can lead to nephrotoxicity, in addition to potassium and magnesium wasting. Flucytosine can also cause neutropenia, especially in patients with chronic kidney disease. Therefore, it is extremely important to monitor electrolyte levels and complete blood cell counts closely. In addition, patients should be monitored clinically for infusion-related reactions during and following each administration. Lumbar puncture is repeated 2 weeks after induction therapy in immunocompromised hosts to assess response to therapy by the presence or absence of fungal growth. It is important to recognize CM at an early stage given its high morbidity and mortality if adequate treatment is not started promptly. The HIV-negative status may lead to a delay in diagnosis, so it is helpful to consider other risk factors that impair cell-mediated immunity, especially prednisone use, which in turn predisposes patients to CM.

The association of Human Herpes virus simplex 1 and 2 infection of male genital tract and male infertility

Objectives: To investigate the association of viral infections present in the lower genital tract of males and relationship among male infertility.Methods: ELISA and RT-PCR technique of 100 semen and serum specimens, collected over 8 months from males investigated for infertility, were retrospectively assessed, by screening anti-human Herpes Simplex Virus (HSV) IgM and IgG in the serum and detection of HSV DNA in seminal fluid.

頭頂葉皮質病変で発症し，当初に脳梗塞との鑑別を要した単純ヘルペス脳炎の1例

A 73-year-old woman was admitted to our hospital due to a decreased conscious level and a high fever. Six days before her admission, she felt transient numbness in her right lower limb. Brain MRI taken by her local doctor revealed only right parietal cortex lesions. She was diagnosed with transient ischemic attack and started on anti-platelet therapy. One day before her admission, she became drowsy, and left-side weakness developed. She was admitted to a community hospital for treating stroke. On the next day, she was referred to our hospital because of a high fever. Our brain MRI showed new lesions in her right temporal lobe. She had no stroke risk factors, and embolic sources were not detected. Cerebrospinal fluid analysis detected herpes simplex virus DNA. She was diagnosed with herpes simplex encephalitis (HSE). HSE is common encephalitis which develops fever, headache and alteration in mental status. It often involves temporal lobe, but extratemporal lesions alone are not uncommon. Diffusion-weighted images (DWI) of brain are of importance to differentiate HSE from stroke. When it is questionable to diagnose with stroke for patients with cerebral cortex lesions, they must be monitored with close observation. There is the possibility of initial presentation of HSE in that situation even if patients have no typical symptoms.

Prevalence of human papillomavirus and herpes simplex virus in amniotic fluid from pregnant women of Eastern Poland.

The exact route of human papillomavirus (HPV) and herpes simplex virus (HSV) transmission from a pregnant woman to her fetus has not been clearly established thus far. The data of many studies raise the possibility of intrauterine infection. In order to clarify our knowledge about virus vertical transmission in pregnant women, viral prevalence in amniotic fluid cannot. To the best of our knowledge, this is the first study on HPV DNA and HSV DNA detection in amniotic fluid in Poland. The study covered 138 samples of amniotic fluid from patients undergoing invasive prenatal diagnostic procedures (for medical indications) during the second trimester of gestation. The aim of the study was to assess the prevalence of HPV and HSV in the amniotic fluid samples obtained from asymptomatic women with intact amniotic membranes. To identify viral DNA of HPV and HSV in collected material, polymerase chain reaction (PCR) was performed. We did not find HPV or HSV DNA in any of the examined specimens of amniotic fluid. Our investigation did not confirm the prenatal transmission of HPV and HSV to the amniotic fluid.

Early diagnosis of herpes encephalitis saves lives: A case report

Herpes simplex virus (HSV) is the most important pathogen causing endemic encephalitis. HSV-1 is responsible for 90% of the cases which is the only treatable viral encephalitis. Clinically it is characterized by decreased level of consciousness, changes in personality, convulsions, and focal neurological deficits. Diagnosis of HSV encephalitis is made by detection of HSV DNA in the cerebrospinal fluid via polymerase chain reaction (PCR) method. The mortality rate in untreated HSV encephalitis is 50-70%. In this paper a patient diagnosed as Herpes simplex encephalitis according to positive PCR, clinic findings, radiological imaging and electroencephalogram (EEG). The patient was successfully treated for 21 days with acyclovir without any complication. This case report emphasizes the value of early diagnosis and treatment.

Development history of herpes simplex encephalitis

Herpes simplex encephalitis (HSE) is an acute central nervous system infection caused by herpes simplex virus (HSV). Early clinical manifestations mainly include fever, headache and unconsciousness; when progressing, psychiatric symptoms can occur. Death or serious neurological sequelae will happen if not treated. With the development of laboratory tests and imaging techniques, the early diagnosis of HSE is possible. Even though imaging with temporal lobe abnormal signal has the implication to HSE, the application of polymerase chain reaction (PCR) in detecting HSV DNA in cerebrospinal fluid is currently the gold standard to diagnose HSE. Once diagnosed, acyclovir must be given as soon as possible, as delayed treatment will result in a poor outcome. doi: 10.3969/j.issn.1672-6731.2014.08.003

Herpes simplex virus reactivation as a trigger of mucous lesions in pemphigus vulgaris

Although infectious agents have long been implicated in the induction or exacerbation of pemphigus vulgaris (PV), a convincing role for the agent in the aetiology of PV has not been established. To establish the association with PV and herpes simplex virus (HSV). We examined saliva for the presence of HSV DNA after the onset of PV initially localized to the oral lesions in addition to conventional serological tests and immunohistochemistry. We successfully detected high levels of HSV DNA in the saliva samples from six of 16 patients with PV at the earliest stage, who had no episodes of herpes simplex. The prevalence (37·5%) of detecting HSV DNA in the patients with PV was lower than that of eczema herpeticum (56·5%), but comparable to that in patients with herpes labialis (30·0%). Copy numbers of the HSV DNA were rather higher than those with herpes labialis and with eczema herpeticum. In general, detection of HSV DNA in saliva was transient and restricted to the earliest phase of the disease. In addition, anti-HSV immunoglobulin (Ig) G titres in patients with PV were significantly higher than those in patients with virologically confirmed HSV-induced disorders. All salivary HSV DNA-positive patients with PV had run a more complex, intractable course refractory to conventional therapy. Detection of HSV DNA in saliva is a useful and noninvasive, quantitative method for establishing the role of HSV in the pathogenesis of PV and for identifying individuals at greater risk for subsequently developing refractory PV.

Neonatal herpes in Serbia: Is it a problem or not?

With 20-80% mortality, neonatal infection caused by herpes simplex virus (HSV) or neonatal herpes is among the most severe of all perinatal infections. The majority of neonatal HSV infections are acquired during delivery, although in utero and postnatal infections do occur. Primary maternal infection is associated with a high rate of transmission (~50%), compared to <3% in infants of women with reactivated disease. Other factors that influence transmission include HSV type, premature delivery, etc. Clinical manifestations have been classified into three forms: skin-eye-mouth disease, CNS and disseminated disease. The diagnosis of neonatal HSV infection includes the detection of HSV DNA by PCR in samples from neonate and mother. The incidence of neonatal herpes differs widely between different countries. In Serbia, the data about neonatal herpes incidence are scarce. The results of our pilot study showed that the minimal estimation of the national incidence of neonatal herpes is 7.5 per 100 000. Therefore, the set up and implementation of a national neonatal herpes surveillance system might provide valuable information for the accurate assessment of disease burden and development of an effective prevention strategy in Serbia.

Encefalitis herpética neonatal: dos gemelas, dos casos

Herpes simplex encephalitis is an infrequent infection with high mortality and morbidity. Antiviral therapies decrease mortality but long term sequelae are still high, so early diagnosis is important for opportune treatment. We present a pair of twins with central nervous system herpes simplex infection during the first month of life. Both twins presented non-specific symptoms and consulted with 48 hours apart needing intensive care admission, the first one for noninvasive mechanical ventilation and the second for hemodynamic support. Diagnosis was made by cerebrospinal fluid PCR, in the first twin at day 9 of disease and in the second at admission. Both twins were treated with acyclovir, but only the second one at the beginning of her illness. Initial study with electroencephalogram and magnetic resonance was normal and cerebrospinal fluid on day 18 of treatment was negative for herpes simplex virus DNA detection in both patients.

Herpes Simplex Encephalitis: From Virus to Therapy

Herpes simplex virus (HSV) is the cause of herpes simplex encephalitis (HSE), a devastating human disease which occurs in 2-4 cases per million/year. HSE results either from a primary infection or virus reactivation, in accordance with the common pattern of HSV infection which is a chronic lifelong process. However its pathophysiology remains largely unknown and its poor prognosis is in contrast with the usually good tolerance of most clinical herpetic manifestations. HSE is due to HSV type 1 (HSV-1) in most cases but HSV type 2 (HSV-2) may be also implicated, especially in infants in the context of neonatal herpes. Polymerase chain reaction detection of HSV DNA in cerebrospinal fluid is the diagnosis of choice for HSE. Acyclovir, a nucleoside analogue which inhibits viral DNA polymerase activity, is the reference treatment of HSE while foscarnet constitutes an alternative therapy and the efficacy of cidofovir is currently uncertain in that context. The emergence of HSV resistance to acyclovir, a phenomenon which is mainly observed among immunocompromised patients, is a current concern although no case of HSE due to an acyclovir-resistant HSV strain has been reported to date. Nevertheless the identification and development of novel therapeutic strategies against HSV appears to be a non dispensable objective for future research in virology.

Diagnosis and quantitative detection of HSV DNA in samples from patients with suspected herpes simplex encephalitis

Diagnosis of herpes simplex encephalitis (HSE) is based on the detection of herpes simplex virus (HSV) DNA in patients’ CSF samples. HSV DNA quantitation has the potential for estimating the effects of antiviral therapy. The aim of this study was to diagnose HSV DNA in HSE suspected patients and the quantitative analysis of its genome using real-time PCR to assess the value of the viral load in the course of antiviral treatment. The CSF samples were collected from 236 consecutive HSE suspected patients from November 2004 to May 2008. Upon DNA extraction, the samples were analyzed by Real-Time PCR assay. A set of primers amplified a common sequence of HSV glycoprotein B gene. The copy numbers of unknown samples were expressed via a standard curve drawn with a known amount of amplified cloned plasmid. Of the 236 samples, 137 (58%) came from males and 99 (42%) from females. The HSV genome was detected in 22 (9.3%) patients by PCR, 13 males/9 females. Serial CSF samples were available from 10 of the 22 patients. The range of the HSV DNA copy numbers in the clinical samples ranged from 2.5 × 102 to 1.7 × 106 copies/mL of CSF. Quantitative PCR results can be helpful in evaluating the efficacy of antiviral therapy in the above-mentioned patients. There is an association between the initial viral load and the duration of treatment course.

Investigation of bacterial and viral agents and immune status in Behcet’s disease patients from Iran

Behçet's disease (BD) is an autoimmune disorder associated with HLA-B51 positivity. Serologic/genomic findings have suggested microbes as possible causative agents and the geographical distribution suggests environmental influences. We performed comparative analyses of 40 patients with BD or related symptoms not fulfilling BD criteria. Patients originating from different regions of Iran were tested by molecular/serological methods for human herpes viruses and parvovirus B19, two Chlamydiae species, as well as Coxiella, Listeria, Yersinia, Leptospira and Mycobacterium paratuberculosis. Human leukocyte antigen-typing was performed: testing of cytokine profiles and immune mediators representative for the cellular immune system, including neopterin/kynurenine production. No apparent differences in interleukin (IL)-4, 6, 8 and 10 were observed, whereas production of soluble IL-2-receptor and tumor necrosis factor (TNF)-alpha were more pronounced in the BD group. Neopterin/kynurenine production was comparable, although both groups showed twice the levels of healthy people. No significant differences of herpes simplex virus (HSV) antibody titres were observed but higher titres against Chlamydophila pneumoniae were found in the controls. In 20 BD patients and controls neither parvovirus B19 DNA was detected nor bacterial DNA. Viral DNA of Epstein-Barr virus (EBV), cytomegalovirus (CMV) and human herpes virus (HHV)8 was detected more frequently in the BD group, whereas HSV DNA was only found in the controls, indicating that stomatitis might be caused by HSV. Although no significant association of BD was detected with a single pathogen, our findings suggest that detection of HSV DNA or Chlamydiae would rather argue against classic BD. Whether there is a discriminative potential of the tested immune mediators/receptors has to be elucidated in further studies.

Correlates of HIV-1 Genital Shedding in Tanzanian Women

BackgroundUnderstanding the correlates of HIV shedding is important to inform strategies to reduce HIV infectiousness. We examined correlates of genital HIV-1 RNA in women who were seropositive for both herpes simplex virus (HSV)-2 and HIV-1 and who were enrolled in a randomised controlled trial of HSV suppressive therapy (aciclovir 400 mg b.i.d vs. placebo) in Tanzania.MethodologySamples, including a cervico-vaginal lavage, were collected and tested for genital HIV-1 and HSV and reproductive tract infections (RTIs) at randomisation and 6, 12 and 24 months follow-up. Data from all women at randomisation and women in the placebo arm during follow-up were analysed using generalised estimating equations to determine the correlates of cervico-vaginal HIV-1 RNA detection and load.Principal FindingsCervico-vaginal HIV-1 RNA was detected at 52.0% of 971 visits among 482 women, and was independently associated with plasma viral load, presence of genital ulcers, pregnancy, bloody cervical or vaginal discharge, abnormal vaginal discharge, cervical ectopy, Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, an intermediate bacterial vaginosis score and HSV DNA detection. Similar factors were associated with genital HIV-1 RNA load.ConclusionsRTIs were associated with increased presence and quantity of genital HIV-1 RNA in this population. These results highlight the importance of integrating effective RTI treatment into HIV care services.

Genital Herpes / Genitalni herpes

Abstract Genital herpes is a chronic, nearly always active herpes simplex virus (HSV) infection of sacral ganglia, that may appear bilaterally and in more ganglia than previously thought. It represents one of the most prevalent sexually transmitted infections, and the most frequent cause of genital ulcer disease in the general populations of developed countries. It is caused by HSV type-2 (HSV-2) in 60-80% of cases, with HSV-1 infections causing the remainder. Genital herpes caused by HSV-1 is on the rise. Since genital HSV-1 infections have higher risk for transmission from mother to infant during delivery than HSV-2, they account for 30% of all cases of neonatal herpes. Serological studies have found prevalence of HSV-2 in the general population of developed contries to be up to 25%. Thirty years ago, herpes was defined as “Today’s Scarlet Letter”in the absence of reliable serological tests and highly effective medications, for diagnosis and treatment of genital herpes. In 2000, apart from virus isolation in cell culture (70% sensitivity), that has long been regarded as the diagnostic gold standard, type specific serological tests and higly effective antiviral agents have evolved. However, the following questions were raised: should serological testing be routinely recommended in asymptomatic patients; can antiviral therapy reduce asymptomatic shedding of the virus; can antiviral therapy reduce sexual transmission of infection; can antiviral therapy reduce acquisitation of viral copathogens, such as human immunodeficiency virus (HIV)? Now, ten years later, we know the answers. Type specific HSV DNA detection by real-time PCR assays (100% sensitivity) are tests of choice for every person with recurrent genital ulcers lasting more than 4 days, and must be available in those laboratories currently performing a significant number of PCR tests for different purposes. Type specific IgG serology assays are indicated in all asymptomatic persons who are at increased risk for HSV infection. In sexually active patients experiencing ≥ 6 recurrences per year, daily supressive dose of acyclovir, valacyclovir or famciclovir should be discontinued after a maximum of a year of continuous antiviral therapy in order to reassess recurrence frequency. If necessary, the therapy should be restarted after at least two recurrences. With such expansive diagnostic possibilities and more aggressive therapeutic approaches, we can define genital herpes not as a “Scarlet Letter”, but as a “widespread untoward consequence of human sexuality”.

Incidence and impact of herpes simplex and cytomegalovirus detection in the respiratory tract after lung transplantation

Herpesvirus infections cause morbidity in lung transplant recipients. The study was conducted to investigate the incidence and impact of herpes simplex virus (HSV) and cytomegalovirus (CMV) detection in the respiratory tract (RT) of lung and heart-lung transplant recipients (LTR) during the postoperative phase. In a prospective cohort study, 91 LTR having at least 1 nasopharyngeal swab (NPS) sent for virus diagnostics were monitored for CMV and HSV detection in NPS during their post-transplant hospital stay on cardiothoracic surgery wards (median 4 weeks) by direct immunofluorescence testing for HSV, virus culture, and CMV and HSV polymerase chain reaction (PCR). Bronchoalveolar lavages (BALs) were analyzed with the same protocol except that HSV PCR was only performed on request. Risk factor analysis for the outcome '90-day mortality' was performed. Fifteen LTR had virus detection in NPS (16.5%): 9 had CMV, 5 had HSV, and 1 had both CMV and HSV. Four of 84 LTR had CMV detection in BAL (4.8%). Absence of CMV detection in NPS had a negative predictive value of 98.8% for absence of CMV detection in BAL. HSV DNA detection in NPS, especially if detected within 8 days after transplantation, was associated with 90-day mortality. In conclusion, detection of herpesviruses in the RT was clinically relevant and frequent, despite antiviral prophylaxis.

Long‐Term Impact of Acyclovir Suppressive Therapy on Genital and Plasma HIV RNA in Tanzanian Women: A Randomized Controlled Trial

Herpes simplex virus (HSV) suppressive therapy reduces genital and plasma human immunodeficiency virus type 1 (HIV-1) RNA over periods up to 3 months, but the long-term effect is unknown. A total of 484 HIV-1 and HSV type 2 seropositive Tanzanian women aged 16-35 years were enrolled in a randomized placebo-controlled trial of acyclovir administered at a dosage of 400 mg twice daily. Cervico-vaginal lavage and blood samples were collected at 6 months, 12 months, and 24 months for quantification of genital and plasma HIV-1 RNA and genital HSV DNA. Primary outcomes were detection and quantity of cervico-vaginal HIV-1 RNA at 6 months. At 6 months, there was little difference between the acyclovir and placebo arms for cervico-vaginal HIV-1 RNA detection (88 [ 41 .3%] of 213 vs 84 [ 44 .0%] of 191; odds ratio [OR], 0.90; 95% confidence interval [CI], 0.60-1.33), HSV DNA detection (20 [ 9 .4%] of 213 vs 22 [ 11 .5%] of 191; OR, 0.80; 95% CI, 0.42-1.51), genital HIV or HSV loads, or plasma HIV-1 RNA load. Estimated median adherence was 91%. There was a suggestion of an impact on cervico-vaginal HIV-1 RNA detection among women with estimated adherence 90% (OR, 0.74; 95% CI, 0.50-1.09) when data from all 3 visits were included. Acyclovir administered at a dosage of 400 mg twice daily is unlikely to be a useful long-term intervention to reduce HIV transmission. The lack of effect on HIV may be attributable to suboptimal adherence or treatment regimen.